Improved supporting technologies for imaging of molecular and supramolecular structures within cells are needed to facilitate cell biology research, and are sought by the National Institute of General Medical Sciences (specifically, its Division of Cell Biology and Biophysics). Multiscale imaging, using cryo-electron tomography (cryo-ET) on supramolecular structures and single molecules, has proven in recent years to be a unique and invaluable method for highthroughput characterization of the dynamic 3D architecture of cells. Electron microscopy (EM) grids, used as substrates for supporting the biological and biomolecular specimens being imaged, are a critical component associated with this imaging method. New EM grid technology that decreases sample preparation cost and time, improves sample generation from culturing to freezing for cryo-ET, and increases imaging quality will allow researchers to more efficiently explore cellular architecture, at higher throughput. Synkera proposes a novel class of ceramic EM grids that feature an integrated thin support film that is highly compatible with cell culturing, light microscopy and cryo-ET. The grids will facilitate high-throughput, multiscale imaging of sub-cellular architecture and offer key advantages over state-of-the art products. The grids are also expected to be a competitive alternative in many other EM and culturing applications a 6-month Phase I project successfully demonstrated feasibility of the proposed grids architecture, as well as their potential in culturing, cryo-ET and multiscale imaging. Phase II aims to build on this success by further developing fabrication processes and ceramic EM grids designs, to fully realize the potential for multiscale imaging in functional prototypes. At least four academic partners will aide in this development process. The ultimate goal of the proposed project is a complete line of ceramic EM grids for a broad range of EM applications, from bioimaging to materials characterization. PUBLIC HEALTH RELEVANCE: The project addresses imaging of molecules and cells via cryo-electron tomography (cryo-ET). Specifically, the target application is multiscale imaging via optical microscopy and cryo-ET of cellular, supramolecular and single-molecule structures, for generating 3D models of sub-cellular architecture. The development of a novel class of ceramic-based electron microscopy grids that facilitate this multiscale imaging is proposed. The proposed technology will offer greater capability over state-of-the- art products and help further streamline multiscale cellular imaging by simplifying the specimen preparation process and yielding superior imaging performance.